Microtechnique in histology, histology sample preparation methods

Introduction for microtechnique.
Histopathology specimen preparation

1. Introduction Dr/ Muaid Albasheri Histopathology and Cytology Lab Founder and Manager National Oncology Center-Aden 01/10/2024 10:13 م 1

2. The study of the structure of tissue by means of special stains technique combined with light microscope /E.M. What is Histology? 01/10/2024 10:13 م 2

3. Why do we study it? Study microscopic structure of cells & tissues Correlate this structure to function Understand the pathology of diseases 01/10/2024 10:13 م 3

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6. 1-preparing thin section 01/10/2024 10:13 م 6

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9. Excisional biopsy : A whole organ or a whole lump is removed ( excised). Incisional biopsy: only portion of the lump is removed( soft tissue). Endoscopic Biopsy :   are used to reach tissue inside the body in order to gather samples from places like the bladder, colon, or lung Colposcopic biopsy: a small sample of tissue (biopsy) may be taken from the cervix or from inside the opening of the cervix ( endocervical canal) 01/10/2024 10:13 م 9 Types of biopsy

10. 1- In the fields of histology , pathology , and cell biology , fixation is a chemical process by which biological tissues are preserved from autolysis or putrefaction . 2-FIXATIVE : A chemical agent used for the preservation and stabilization of tissues for microscopical study . 01/10/2024 10:13 م 10 Fixation and Fixative

11. Functions and characteristics of a good fixative - It hardens the tissue by coagulating its protein. - It prevents putrefaction and stops the autolytic changes by killing the bacteria. - It preserves the tissue in a condition similar to that existing during life. - It facilitates the process of cutting, staining and microscopic examination . 01/10/2024 10:13 م 11

12. Automatic tissue processor 01/10/2024 10:13 م 12

13. 3-dehydration 01/10/2024 10:13 م 13

14. 4-clearing 01/10/2024 10:13 م 14

15. -59c 5-Impregnation Infiltration with Paraffin wax in 55. Purpose of paraffin wax infilteration : 1-Smoother handle 2-Damage free-cutting of tissue 2-Aid in firm consistency 01/10/2024 10:13 م 15

16. The aim of tissue processing is to embed the tissue in a solid medium firm enough to support the tissue and enable thin sections to be cut , and yet soft enough not to damage the knife or tissue. Aim of tissue processing 01/10/2024 10:13 م 16

17. 6-Embedding in paraffin wax 01/10/2024 10:13 م 17

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19. 8 -Floating and Mounting 01/10/2024 10:13 م 19

20. Paraffin technique. 01/10/2024 10:13 م 20

21. II.STAINING 01/10/2024 10:13 م 21

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24. Steps for Staining with Haematoxylin and Eosin (H & E) 1- Deparaffinize in xylene (5 min). 2- Hydrate in descending grades of alcohol 3- Stain in Hematoxylen for 2 min. 4- Bluing section in tap water 5- Stain in eosin for 30 second. 6- Wash in distilled water. 7- Dehydrate in ascending grades of alcohol. 8- Clear in xylene. 9- Mount in D.P.X & cover with a cover slip to protect and fix the section on the slide . 01/10/2024 10:13 م 24

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30. Thank you for attention 01/10/2024 10:13 م 30